Cell Type-Specific Oxidative Stress Genomic Signatures in the Globus Pallidus of Dopamine-Depleted Mice.
Cell Type-Specific Oxidative Stress Genomic Signatures in the Globus Pallidus of Dopamine-Depleted Mice.
复制标题
多巴胺耗竭小鼠苍白球中细胞类型特异性氧化应激基因组特征。
DOI：
10.1523/jneurosci.1634-20.2020
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发表时间：
2020-12-09
期刊：
影响因子：
--
通讯作者：
Pfenning AR
中科院分区：
文献类型：
--
作者：
Lawler AJ;Brown AR;Bouchard RS;Toong N;Kim Y;Velraj N;Fox G;Kleyman M;Kang B;Gittis AH;Pfenning AR
神经元亚型功能障碍是神经系统疾病回路的一个关键因素，但识别相关的基因调控途径因大脑的分子复杂性而变得复杂。例如，外苍白球（GPe）中的小清蛋白表达（PV+）神经元与帕金森病的多巴胺耗尽小鼠模型的运动缺陷密切相关，其中PV+神经元的细胞类型特异性光遗传学刺激超过其他神经元群体拯救了运动。尽管这些细胞类型在神经回路中发挥着不同的作用，但由于难以分离罕见的神经元亚型，因此分子相关性仍然未知。为了解决这个问题，我们开发了一种新的病毒亲和纯化策略，Cre特异性核锚定独立标记，从成年小鼠脑中分离Cre重组酶表达（Cre+）核。应用该技术，我们对雄性和雌性小鼠的三个脑区（GPe、纹状体和皮质）内多巴胺耗竭对PV+和PV-细胞的细胞类型特异性转录组学和表观遗传学影响进行了靶向评估。我们发现GPe PV+神经元特异性基因表达的变化表明缺氧诱导因子2α信号的增加。与转录组学数据一致，受GPe PV+神经元内多巴胺耗竭影响的开放染色质区域富含缺氧诱导因子家族结合基序。对通过Cre-Specific Nuclear Anchored Independent Labeling分离的PV+神经元进行的基因表达和表观基因组实验发现，由神经保护因子Hif 2a介导的转录调节网络是响应多巴胺耗尽的潜在神经回路差异。重要性声明Cre-Specific Nuclear Anchored Independent Labeling是一种增强的基于病毒的方法，用于分离特定细胞类型的细胞核，用于转录组和表观基因组研究，减少对转基因动物的依赖。将这项技术应用于帕金森病小鼠模型中表达GPe小白蛋白的神经元，我们发现了涉及缺氧诱导因子2α的氧稳态维持途径上调的证据。这些结果提供了新的见解，黑质外的神经元亚型可能在分子水平上补偿帕金森病进展过程中运动产生神经回路的差异。此外，他们强调了细胞类型特异性技术的实用性，例如Cre-Specific Nuclear Anchored Independent Labeling，用于复杂系统中特定神经元亚型的隔离评估。
Neuron subtype dysfunction is a key contributor to neurologic disease circuits, but identifying associated gene regulatory pathways is complicated by the molecular complexity of the brain. For example, parvalbumin-expressing (PV+) neurons in the external globus pallidus (GPe) are critically involved in the motor deficits of dopamine-depleted mouse models of Parkinson's disease, where cell type-specific optogenetic stimulation of PV+ neurons over other neuron populations rescues locomotion. Despite the distinct roles these cell types play in the neural circuit, the molecular correlates remain unknown because of the difficulty of isolating rare neuron subtypes. To address this issue, we developed a new viral affinity purification strategy, Cre-Specific Nuclear Anchored Independent Labeling, to isolate Cre recombinase-expressing (Cre+) nuclei from the adult mouse brain. Applying this technology, we performed targeted assessments of the cell type-specific transcriptomic and epigenetic effects of dopamine depletion on PV+ and PV– cells within three brain regions of male and female mice: GPe, striatum, and cortex. We found GPe PV+ neuron-specific gene expression changes that suggested increased hypoxia-inducible factor 2α signaling. Consistent with transcriptomic data, regions of open chromatin affected by dopamine depletion within GPe PV+ neurons were enriched for hypoxia-inducible factor family binding motifs. The gene expression and epigenomic experiments performed on PV+ neurons isolated by Cre-Specific Nuclear Anchored Independent Labeling identified a transcriptional regulatory network mediated by the neuroprotective factor Hif2a as underlying neural circuit differences in response to dopamine depletion. SIGNIFICANCE STATEMENT Cre-Specific Nuclear Anchored Independent Labeling is an enhanced, virus-based approach to isolate nuclei of a specific cell type for transcriptome and epigenome interrogation that decreases dependency on transgenic animals. Applying this technology to GPe parvalbumin-expressing neurons in a mouse model of Parkinson's disease, we discovered evidence for an upregulation of the oxygen homeostasis maintaining pathway involving Hypoxia-inducible factor 2α. These results provide new insight into how neuron subtypes outside the substantia nigra pars compacta may be compensating at a molecular level for differences in the motor production neural circuit during the progression of Parkinson's disease. Furthermore, they emphasize the utility of cell type-specific technologies, such as Cre-Specific Nuclear Anchored Independent Labeling, for isolated assessment of specific neuron subtypes in complex systems.
影响因子：
2.7
作者：
Day, Jeremy J;Sweatt, J David
通讯作者：
Sweatt, J David